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Matrix attachment region from the chicken lysozyme locus reduces variability in transgene expression and confers copy number-dependence in transgenic rice plants

  • Genetic Transformation and Hybridization
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Abstract

Matrix-attachment regions (MARs) may function as domain boundaries and partition chromosomes into independently regulated units. In this study, BP-MAR, a 1.3-kb upstream fragment of the 5′MAR flanking the chicken lysozyme locus, was tested for its effects on integration and expression of transgenes in transgenic rice plants. Using the Agrobacterium-mediated method, we transformed rice with nine different constructs containing seven and six different promoters and coding sequences, respectively. Genomic Southern blot analyses of 357 independent transgenic lines revealed that in the presence of BP-MAR, 57% of the lines contained a single copy of the transgene, whereas in its absence, only 20% of the lines contained a single copy of the transgene. RNA gel-blot and immunoblot experiments demonstrated that in the presence of BP-MAR, transgene expression levels were similar among different lines. These data were in direct contrast to those derived from transgenes expressed in the absence of BP-MAR, which varied markedly with the chromosomal integration site . Thus, it can be concluded that BP-MAR significantly reduces the variability in transgene expression between independent transformants. Moreover, the presence of BP-MAR appears to confer a copy number-dependent increase in transgene expression, although it does not increase expression levels of individual transgenes. These data contrast with results previously obtained with various MARs that increased expression levels of transgene significantly. Therefore, we conclude that the incorporation of BP-MAR sequences into the design of transformation vectors can minimize position effects and regulate transgene expression in a copy number-dependent way.

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Acknowledgements

We are grateful to Dr. Jan-Peter Nap at Wageningen University and Research Center for providing pUC-B1-X1. This work was supported by the Ministry of Science and Technology through the Crop Functional Genomics Center (grants to J.-K.K. and B.H.N.), by the Rural Development Administration through the Biogreen21 Program (grant to J.-K.K.), by the Korea Science and Engineering Foundation through the Plant Metabolism Research Center at Kyung-Hee University (grant to J.-K.K.) and by the Ministry of Agriculture (grant to J.-K.K.).

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Authors and Affiliations

  1. Division of Bioscience and Bioinformatics, Myongji University, Yongin, 449-728, Korea

    S.-J. Oh, J. S. Jeong, E.-H. Kim, I.-C. Jang, B. H. Nahm & J.-K. Kim

  2. Genomics and Genetics Institute, GreenGene Biotech, Yongin, 449-728, Korea

    N. R. Yi, Y. S. Kim & B. H. Nahm

  3. National Seed Management Office, Suwon, 440-310, Korea

    S.-I. Yi

  4. National Institute of Agricultural Biotechnology, RDA, Suwon, 440-770, Korea

    S.-C. Suh

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  1. S.-J. Oh
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  2. J. S. Jeong
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  3. E.-H. Kim
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  4. N. R. Yi
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  5. S.-I. Yi
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  6. I.-C. Jang
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  7. Y. S. Kim
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  8. S.-C. Suh
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  9. B. H. Nahm
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  10. J.-K. Kim
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Correspondence to J.-K. Kim.

Additional information

Communicated by I.S. Chung

S.-J. Oh, J.S. Jeong, E.-H. Kim, N.R. Yi and S.-I. Yi contributed equally to the paper

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Oh, SJ., Jeong, J.S., Kim, EH. et al. Matrix attachment region from the chicken lysozyme locus reduces variability in transgene expression and confers copy number-dependence in transgenic rice plants. Plant Cell Rep 24, 145–154 (2005). https://doi.org/10.1007/s00299-005-0915-2

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  • DOI: https://doi.org/10.1007/s00299-005-0915-2

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